Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering.

2.50
Hdl Handle:
http://hdl.handle.net/10033/601366
Title:
Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering.
Authors:
Tu, Qiang; Herrmann, Jennifer; Hu, Shengbiao; Raju, Ritesh; Bian, Xiaoying; Zhang, Youming; Müller, Rolf ( 0000-0002-1042-5665 )
Abstract:
Disorazol, a macrocyclic polykitide produced by the myxobacterium Sorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) from Sorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthus DK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression in M. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.
Affiliation:
Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany.
Citation:
Genetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering. 2016, 6:21066 Sci Rep
Journal:
Scientific reports
Issue Date:
2016
URI:
http://hdl.handle.net/10033/601366
DOI:
10.1038/srep21066
PubMed ID:
26875499
Type:
Article
Language:
en
ISSN:
2045-2322
Appears in Collections:
publications of the department of microbial natural substances ([HIPS]MINS)

Full metadata record

DC FieldValue Language
dc.contributor.authorTu, Qiangen
dc.contributor.authorHerrmann, Jenniferen
dc.contributor.authorHu, Shengbiaoen
dc.contributor.authorRaju, Riteshen
dc.contributor.authorBian, Xiaoyingen
dc.contributor.authorZhang, Youmingen
dc.contributor.authorMüller, Rolfen
dc.date.accessioned2016-03-15T15:07:40Zen
dc.date.available2016-03-15T15:07:40Zen
dc.date.issued2016en
dc.identifier.citationGenetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering. 2016, 6:21066 Sci Repen
dc.identifier.issn2045-2322en
dc.identifier.pmid26875499en
dc.identifier.doi10.1038/srep21066en
dc.identifier.urihttp://hdl.handle.net/10033/601366en
dc.description.abstractDisorazol, a macrocyclic polykitide produced by the myxobacterium Sorangium cellulosum So ce12 and it is reported to have potential cytotoxic activity towards several cancer cell lines, including multi-drug resistant cells. The disorazol biosynthetic gene cluster (dis) from Sorangium cellulosum (So ce12) was identified by transposon mutagenesis and cloned in a bacterial artificial chromosome (BAC) library. The 58-kb dis core gene cluster was reconstituted from BACs via Red/ET recombineering and expressed in Myxococcus xanthus DK1622. For the first time ever, a myxobacterial trans-AT polyketide synthase has been expressed heterologously in this study. Expression in M. xanthus allowed us to optimize the yield of several biosynthetic products using promoter engineering. The insertion of an artificial synthetic promoter upstream of the disD gene encoding a discrete acyl transferase (AT), together with an oxidoreductase (Or), resulted in 7-fold increase in disorazol production. The successful reconstitution and expression of the genetic sequences encoding for these promising cytotoxic compounds will allow combinatorial biosynthesis to generate novel disorazol derivatives for further bioactivity evaluation.en
dc.language.isoenen
dc.titleGenetic engineering and heterologous expression of the disorazol biosynthetic gene cluster via Red/ET recombineering.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Institute for Pharmaceutical Research Saarland (HIPS),Saarland 9 University, 66123 Saarbrücken, Germany.en
dc.identifier.journalScientific reportsen

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